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Hydroxyl Radical Scavenging Activity

The document outlines a method for assessing hydroxyl radical scavenging activity using an ascorbic acid-iron-EDTA model. It details the preparation of various reagents and the procedure for conducting the assay with different concentrations of plant extracts. The intensity of the resulting color is measured to calculate the percentage of scavenging activity.

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7 views1 page

Hydroxyl Radical Scavenging Activity

The document outlines a method for assessing hydroxyl radical scavenging activity using an ascorbic acid-iron-EDTA model. It details the preparation of various reagents and the procedure for conducting the assay with different concentrations of plant extracts. The intensity of the resulting color is measured to calculate the percentage of scavenging activity.

Uploaded by

stylusepson9
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Hydroxyl Radical Scavenging Activity

Hydroxyl radical is one of the potent reactive oxygen species in the biological system. It react
with polyunsaturated fatty acids moieties of the cell membrane phospholipids and cause
damage to cell.

Principle:

Hydoxyl radical scavenging assay is used to find the scavenging activity of free hydroxyl
radical in presence of different concentration of plant samples. The model used is ascorbic
acid-iron-EDTA model of hydroxyl radical generating system. This is a totally aqueous
system in which ascorbic acid, iron and EDTA conspire with each other to generate hydroxyl
radical.

Reagent Preparation:

1. Iron-EDTA solution was prepared by mixing 0.13% ferrous ammonium sulphate and
0.26% EDTA.

2. 0.018% EDTA was prepared by dissolving 0.018g EDTA in 100ml distilled water.

3. 0.22% ascorbic acid was prepared by dissolving 0.22g ascorbic acid in 100 ml distilled
water.

4. 17.5% Trichloroacetic acid (TCA) was prepared by dissolving 17.5gm TCA in 100ml of
distilled water.

5. Nash reagent was prepared by adding 7.5gm ammonium acetate, 0.5ml glacial acetic acid
and 0.2ml of acetone to 100ml distilled water.

6. DMSO (0.85% in 0.1M phosphate buffer, pH7.4)

Procedure:

Various concentration of extract (250, 500, 750 and 1000µg/ml) were taken and 1ml of iron-
EDTA solution. 0.5ml EDTA solution, 1ml DMSO and 0.5ml of ascorbic acid were added.
The mixture was incubated in boiling water bath at 80 to 90°C for 15 min. After incubation
1ml of ice cold TCA and 3ml Nash reagent were added and then the reaction mixture were
incubated at room temperature for 15 min for colour development. The intensity of yellow
colour was measured at 412 nm against a reagent blank. The % hydroxyl radical scavenging
activity is calculated by the following formula:

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